The current work aims to investigate the effect of abiotic stresses (nitrogen (N) and sulfur (S) [0.0 g/l, 1.5 g/l, 3 g/l, 6 g/l, and 12 g/l N and 0.0 g/l, 0.07 g/l, 0.15 g/l, 0.3 g/l, and 0.6 g/l S] and their combination [0.3 g/l S + 6 g/l N]) of axenic culture of
Nostoc linckia
on the production of secondary metabolites which induce different biological activities. Growth rate was measured by dry weight (DW) and optical density (OD)
550 nm
. Additionally, phytochemical compounds, defense enzymes as well as antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(ethylbenzthiazoline-6-sulfonic acid) (ABTS
·+
) radical assays of crude extracts (methylene chloride:methanol, 1:1) were evaluated. Based on antioxidant activity, four promising extracts were subjected to different biological activities such as anticancer, antimicrobial, and antiviral activities. The obtained results revealed that supplementation of external nitrogen source in the form of sodium nitrate was found to increase the total phycobiliprotein content by fivefold. Also, nitrogen depletion provoked significantly highest quantities of phenolic and flavonoid content and this has effects on biological activities of
Nostoc linckia
. Moreover, 0.3 g/l S was found to be the most effective extract exhibiting a significant increase in antioxidant activity based on DPPH and ABTS assays, respectively (88.18 ± 0.64% and 84.20 ± 1.01%). Furthermore, it recorded anticancer activity against HCT 116 cell line with IC
50
of 155 μg/ml. Moreover, this extract possessed a noticeable antibacterial potency (21.0 ± 1.0 as mm inhibition zone against
Staphylococcus aureus
and 19.3 ± 0.6 against
Streptococcus mutans
). In addition, its antiviral activity against H
5
N
1
virus as a percentage of inhibition was 50% and 63.6% at a concentration of 7 μg/ml and 28 μg/ml, respectively, with cytotoxicity less than 7 μg/μl. GC-MS analysis recorded the presence of bioactive compounds exhibiting different biological activities. Therefore, the obtained results can represent valuable bioactive compounds with variable biological potencies.